Immunoenzymic studies on testicular hyaluronidase from rhesus monkeys (Macaca mulatta).
Hyaluronidase from rhesus monkey testes was purified by detergent extraction, ammonium sulphate fractionation, Sephadex G-200 column chromatography and concanavalin A-Sepharose affinity chromatography. The purified hyaluronidase showed one protein band on acrylamide gel electrophoresis. Antibodies to the purified hyaluronidase were raised in rabbits and showed a single precipitin line by Ouchterlony gel diffusion. The enzyme had a molecular weight of 62,000. The Km was 0.5 mg/ml for hydrolysis of hyaluronic acid at 37 degrees C. The optimum pH for the enzyme was 5.0 but activity was present over a broad pH range. The hyaluronidase was inhibited by HgCl2, CuSO4, FeSO4 and p-chloromercuribenzoate all at a concentration of 2 x 10(-4) M. Cysteine protected the enzyme against HgCl2 inhibition.
D000818:Animals; D002845:Chromatography; D003429:Cross Reactions; D006821:Hyaluronoglucosaminidase / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D005779:Immunodiffusion; D008253:Macaca mulatta; D008297:Male; D008970:Molecular Weight; D011817:Rabbits; D013737:Testis / Q000201:enzymology*
D006821:Hyaluronoglucosaminidase